Appendix A: To establish and maintain a rodent breeding colony
2015-2016
PI Name (Last name, First name):
Campus Phone Number:
Title of Protocol Associated with this Appendix:
Number of Experimental Protocol
Will You Be Breeding Transgenic Animals?
No
Yes
If No, complete only Section I; If Yes, complete both Sections I & II.
Are You Creating Any New Transgenic Lines?
No
Yes
If Yes, you will need to obtain approval from the Office of Environmental Health and Safety prior to
receiving final approval from the IACUC. A copy of the approval should be attached.
…SECTION 1.
BREEDING PROTOCOL…
A. List the species and strains of animals that will be used for the breeding program.
Expand the table as necessary.
Species
Strain
B. Breeding Colony Justification
Provide the rationale for establishing and maintaining this breeding colony. Include an
explanation as to why animals from commercial sources are not appropriate (e.g., animals are not
commercially available or in very limited supply, reproduction studies, etc.) Note that the cost of
commercially available animals cannot be used as justification.
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C. Husbandry
Unless special arrangements have been made, it is assumed that BRF staff will be in charge of
routine husbandry of the animals. The PI is responsible for:
 Colony management, genotyping, timely weaning of litters, and prevention of
overcrowded cages;
 Ensuring effective communication with husbandry and veterinary staff (e.g. email or
voicemail that is routinely checked);
 Ensuring that overcrowded cages are separated promptly (within the same day as
notified).
 Ensuring adequate space is available prior to setting up new breeding cages or discussing
options with BRF management.
1.
Describe the breeding paradigm and breeding strategy to be used (e.g. pair, harem).
2.
When will the pups be weaned? If NOT consistent with IACUC Guidelines discuss
reasons and details here.
3.
Who will be responsible for monitoring the breeding colony and weaning the animals?
D. Recordkeeping
The PI is responsible for maintaining accurate and clear records of all animals produced and
used. These records must be available for review during laboratory inspections by IACUC.
1.
Briefly describe the record-keeping system that will be used indicating how breeding,
health and maintenance of the colony is recorded
(Please attach a sample record-keeping sheet):
2.
Who will be responsible for maintaining these records?
E. Animal Numbers
NOTE: You may use your own formulas, or worksheets on estimating animal numbers for breeding
colonies provided in: http://www.ncbi.nlm.nih.gov/books/NBK43325/ or the Colony Planning Worksheet
provided by Jackson labs: http://jaxmice.jax.org/support/husbandry/colony-planning.html
1.
Estimate the Number of Animals Needed to Establish and Maintain the Colony
In the table below, estimate the number of breeders necessary to establish and maintain the
colony for the duration of the project (3 years maximum). These are animals held but not
subjected to any experimental manipulation. Be sure to include this number in your animal
protocol.
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Strain/Stock/Genetics
Sex
# Required
2. Estimated Number of Weaned or Adult Animals to be produced and used in
experiments in the approved animal protocol
In the table below, list the number of offspring that will weaned and then subsequently used in
experiments in the attached animal protocol. Be sure to include this number in the animal
protocol, assigning appropriate pain/distress categories.
Strain/Stock/Genetics
Estimate for 1 yr
Estimate for 3 yrs
3. Estimated Number of Suckling Animals (pups not yet weaned) on which some type
of manipulation will be done (e.g. tail-clip or other biopsy for genotyping)
In the table below, estimate the total number of offspring expected from breeding that will be
genotyped. (If not genotyping this # will be zero; note also that if genotyping, not all of the
animals will be used in experiments after weaning).
Strain/Stock/Genetics
Estimate for 1 yr
Estimate for 3 yrs
4. Estimated Number of Suckling Animals to be euthanized at or before weaning
(e.g. excess animals, wrong sex, wrong genotype)
Strain/Stock/Genetics
Estimate for 1 yr
Estimate for 3 yrs
F. Disposition of excess/unsuitable animals
Describe what will be done with any animals that are bred in excess of your needs, or that don't
meet particular genetic or other criteria (e.g. euthanize and dispose). Note that transfer of excess
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animals to another protocol (yours or another investigator) is encouraged and requires completion and
approval of an Animal Transfer form.
G. Euthanasia
Indicate which of the following method(s) will be used (Check all that apply):
*CO2 exposure
Overdose using anesthetic agent
by injection or inhalation *
Cervical dislocation under
general anesthesia
Decapitation under general
anesthesia
Exsanguination under general
anesthesia
*If CO2 or anesthetic drug inhalation will be used, what secondary method(s)
will be used to confirm death? (required)
Decapitation
Cervical Dislocation
Opening Thorax
Exsanquination or Major Organ Removal (e.g. brain, heart)
If anesthetic drugs will be used provide the following details:
Name of agent
Dose (mg/kg or %
concentration)
Route of administration
If anesthetic gases will be used for euthanasia, indicate how these will be scavenged:
Filtering with charcoal canister
Used in a fume hood
Other (describe:
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…SECTION II.
TRANSGENIC ANIMALS…
Complete this section only if you will be breeding transgenic animals (regardless of whether
generated in-house or obtained elsewhere).
A. What DNA/transgene(s) is/are disrupted?
B. How will you monitor for the presence of the transgene(s) in the animals?
Please provide details here (ear punch, tail clip, blood collection, etc.). Note that it is
recommended that tail clip biopsies be done before 17 days of age when the bones are not
ossified. If tail clipping is performed after 21 days of age, it is considered a minor survival
surgery and requires the use of an anesthetic agent as well as subsequent analgesia. This
information must be presented in the animal protocol associated with this breeding appendix.
C. How will genotyped pups be individually identified/ discerned after genotyping has
been performed?
D. What are the expected issues and problems associated with the transgenic line(s)?
Provide a description of any expected clinical signs and describe how the animals will be
monitored and evaluated for possible adverse effects of the genetic manipulation.
E. Provide an estimate of the Pain/Distress levels of the transgenic animals that will be
bred. Note that this classification is not based on anything you will do to the animals, but
rather on the inherent pain and/or distress potentially associated with the genetic alterations that
are created. Use the classification choices listed below the Table.
Transgenic type
(abbreviation)
Estimated # animals
Pain/Distress Category
Classification T-C: No pain/distress is inherent to the transgenic line.
Classification T-D: Pain/distress is expected with the transgenic line and appropriate
analgesia/anesthesia/tranquilizers will be given to alleviate pain/distress.
Classification T-E. Pain/distress is expected given the transgenic line being created and
analgesia/anesthesia/tranquilizers will NOT be given to alleviate pain/distress.
F. What will be done to alleviate pain and/or distress in animals if observed to occur?
Indicate if analgesics will be given (with details) or if euthanasia will be done. If proposing to
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generate category E transgenic animals* you must provide scientific justification as well as
details about animal monitoring.
* This category will require special review by the veterinarian and IACUC.
G. Other: Use this space to provide any additional information or clarifications about the
transgenic animals to be generated that has not been covered previously and that you feel is
important for the IACUC to know.
H. Signatures and Approval
_________________________________________
Signature of Principal Investigator
_________________
DATE
_________________________________________
IACUC Chair Signature
_________________
DATE
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